Methods for reducing contamination in a biological substance

ABSTRACT

Provided herein are materials and methods of reducing contamination in a biological substance or treating contamination in a subject by one or more toxins comprising contacting the biological substance with an effective amount of a sorbent capable of sorbing the toxin, wherein the sorbent comprises a plurality of pores ranging from 50 Å to 40,000 Å with a pore volume of 0.5 cc/g to 5.0 cc/g and a size of 0.05 mm to 2 cm and sorbing the toxin. Also provided are kits to reduce contamination by one or more toxins in a biological substance comprising a sorbent capable of sorbing a toxin, wherein the sorbent comprises a plurality of pores ranging from 50 Å to 40,000 Å with a pore volume of 0.5 cc/g to 5.0 cc/g and a size of 0.05 mm to 2 cm and a vessel to store said sorbent when not in use together with packaging for same.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. patent application Ser. No.14/410,901, filed Dec. 23, 2014, and claims the benefit of PCT PatentApplication No. PCT/US2013/048615, filed Jun. 28, 2013 and U.S.Provisional Application No. 61/666,626, filed Jun. 29, 2012, each ofwhich is incorporated herein by reference in its entirety.

TECHNICAL FIELD

The present invention relates generally to materials, methods, kits anddevices for the reduction of toxin contamination.

BACKGROUND

Toxins are exogenous or endogenous substances that cause a disruption innormal physiologic functions and may cause disease. They cause diseaseby coming in contact with or being absorbed by body tissues that includethe intestines, skin or mucosal membranes. There are thousands, if notmillions of substances that can act as toxins. Even some substances thatare normally not toxins can become toxins under the proper conditions.Toxins vary significantly in their strength and the rapidity in whichthey act. Treatments have been developed for exposure to toxins andinclude removal from contact with the toxin, and the use of antidotes.

Antidotes are substances that can counter act the effects of toxins.However, most antidotes are specific to only one type of toxin or familyof toxins. Therefore, it is impossible for hospitals, clinics, fieldhospitals, doctor's offices, ambulances, and other first responders tocarry an antidote for every toxin. In addition, antidotes have not beendeveloped for many toxins and quantities of some antidotes may be toodistant to effectively be used in the treatment of some exposures totoxins.

Thus, there is a need for new and better materials, methods, kits anddevices for the quick, effective and efficient reduction of toxincontamination and treatment of toxin contamination in a subject.

SUMMARY

Provided herein are suitable materials and methods of reducingcontamination by one or more toxins in a biological substance comprisingcontacting the biological substance with an effective amount of asorbent capable of sorbing the toxin, wherein the sorbent comprises aplurality of pores ranging from 50 Å to 40,000 Å with a pore volume of0.5 cc/g to 5.0 cc/g and a size of 0.05 mm to 2 cm and sorbing thetoxin. Also provided herein are suitable methods of treatingcontamination by one or more toxins in a subject in need thereofcomprising contacting a biological substance of the subject with aneffective amount of a sorbent capable of sorbing the toxin, wherein thesorbent comprises a plurality of pores ranging from 50 Å to 40,000 Åwith a pore volume of 0.5 cc/g to 5.0 cc/g and a size of 0.05 mm to 2 cmand sorbing the toxin.

Provided herein are also suitable kits to reduce contamination by one ormore toxins in a biological substance comprising a sorbent capable ofsorbing a toxin, wherein the sorbent comprises a plurality of poresranging from 50 Å to 40,000 Å with a pore volume of 0.5 cc/g to 5.0 cc/gand a size of 0.05 mm to 2 cm and a vessel to store said sorbent whennot in use together with packaging for same. Also provided herein aresuitable devices to reduce contamination by one or more toxins in abiological substance comprising a sorbent capable of sorbing a toxin,wherein the sorbent comprises a plurality of pores ranging from 50 Å to40,000 Å with a pore volume of 0.5 cc/g to 5.0 cc/g and a size of 0.05mm to 2 cm and a vessel wherein the sorbent is located inside the vesselsuch that the biological substance can be directly introduced into thevessel.

Also provided herein are suitable pharmaceutical compositions comprisinga sorbent capable of sorbing a toxin, wherein the sorbent comprises aplurality of pores ranging from 50 Å to 40,000 Å with a pore volume of0.5 cc/g to 5.0 cc/g and a size of 0.05 mm to 2 cm and a food product ora potable liquid.

Provided herein are methods of reducing contamination by one or moretoxins in a biological substance comprising contacting the biologicalsubstance with an effective amount of a sorbent capable of sorbing oneor more non-toxic subunits, wherein when two or more of those subunitsare combined forms a toxin, wherein the sorbent comprises a plurality ofpores ranging from 50 Å to 40,000 Å with a pore volume of 0.5 cc/g to5.0 cc/g and a size of 0.05 mm to 2 cm, and sorbing the one or morenon-toxic subunits. Also provided herein are methods of treatingcontamination by one or more toxins in a subject in need thereofcomprising contacting a biological substance of the subject with aneffective amount of a sorbent capable of sorbing one or more non-toxicsubunits, wherein when two or more of those subunits are combined formsa toxin, wherein the sorbent comprises a plurality of pores ranging from50 Å to 40,000 Å with a pore volume of 0.5 cc/g to 5.0 cc/g and a sizeof 0.05 mm to 2 cm, and sorbing the one or more non-toxic subunits.

BRIEF DESCRIPTION OF THE DRAWINGS

The summary, as well as the following detailed description, is furtherunderstood when read in conjunction with the appended drawings. For thepurpose of illustrating the invention, there are shown in the drawingsexemplary embodiments of the invention; however, the invention is notlimited to the specific methods, compositions, and devices disclosed. Inaddition, the drawings are not necessarily drawn to scale. In thedrawings:

FIG. 1 illustrates pore volume of the sorbent plotted as a function ofthe pore diameter of the sorbent on a log scale.

FIG. 2 illustrates the Clostridium difficile Toxin A removal as afunction of time.

FIG. 3 illustrates the Clostridium difficile Toxin B removal as afunction of time.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

The present invention may be understood more readily by reference to thefollowing detailed description taken in connection with the accompanyingfigures and examples, which form a part of this disclosure. It is to beunderstood that this invention is not limited to the specific materials,devices, methods, applications, conditions or parameters describedand/or shown herein, and that the terminology used herein is for thepurpose of describing particular embodiments by way of example only andis not intended to be limiting of the claimed invention. The term“plurality”, as used herein, means more than one. When a range of valuesis expressed, another embodiment includes from the one particular valueand/or to the other particular value. Similarly, when values areexpressed as approximations, by use of the antecedent “about,” it willbe understood that the particular value forms another embodiment. Allranges are inclusive and combinable.

It is to be appreciated that certain features of the invention whichare, for clarity, described herein in the context of separateembodiments, may also be provided in combination in a single embodiment.Conversely, various features of the invention that are, for brevity,described in the context of a single embodiment, may also be providedseparately or in any subcombination. Further reference to values statedin ranges include each and every value within that range.

The following definitions are intended to assist in understanding thepresent invention:

The term “anti-microbial agent” includes antibacterial agents,anti-viral agents, antifungal agents, antiseptics and the like. Suitableantimicrobial agents include, but are not limited to isoniazid,rifampin, pyrazinamide, ethambutol, erythromycin, vancomycin,tetracycline, chloramphenicol, sulfonamides, gentamicin, amoxicillin,penicillin, streptomycin, p-aminosalicyclic acid, clarithromycin,clofazimine, minocycline, sulfonamides, ethionamide, cycloserine,kanamycin, amikacin, capreomycin, viomycin, thiacetazone, rifabutin andthe quinolones, such as ciprofloxacin, ofloxacin and sparfloxicin,rifampin, oseltamivir, acyclovir, lamivudine, azole antifungals,echinocandins, and others. Antibacterial agents includes but are notlimited to β-lactam antibacterial agents including, e.g. carbenicillin;ampicillin, cloxacillin, oxacillin and pieracillin, cephalosporins andother cephems including, e.g. cefaclor, cefamandole, cefazolin,cefoperazone, ceftaxime, cefoxitin, ceftazidime, ceftriazone andcarbapenems including, e.g., imipenem and meropenem; and glycopeptides,macrolides, quinolones (e.g. nalidixic acid), tetracyclines, andaminoglycosides (e.g. Gentamicin and Paromomycin).

The term “biocompatible” is defined to mean the sorbent is capable ofcoming in contact with physiologic fluids, living tissues, or organismswithout producing unacceptable clinical changes during the time that thesorbent is in contact with the physiologic fluids, living tissues, ororganisms. In some embodiments, it is intended that the sorbent istolerated by the gut and alimentary canal of the organism. The sorbentsof the present invention are preferably non-toxic. A biocompatiblesorbent may be a biodegradable polymer, a resorbable polymer, or both.

The term “microbe” includes a bacteria, viruses, fungi and parasites.

The term “toxin” is used to mean a substance identified as anetiological agent linked to a negative clinical outcome. Toxins includetoxin subunits, toxin precursors, and virulence factors. The toxin canbe from a biological source such as bacteria, viruses, fungi, parasites,plants or animals. A toxin may be pre-formed or may only become toxiconce in the presence of a biological substance. For example, botulinumtoxin (one of 7 subtypes) is made by the bacterium, Clostridiumbotulinum, often under anaerobic conditions, such as in canned goods.When ingested, this pre-formed botulinum toxin is one of the most potenttoxins known, blocking neuromuscular transmission by inhibitingacetylcholine release in the synapse, causing paralysis and respiratoryfailure. Ricin toxin, made from castor beans, is another example of apre-formed toxin that when inhaled, injected, or ingested can be fatal.Amatoxins, from mushrooms, and aflatoxin, which are mycotoxins producedby strains of the fungus species, Aspergillus, are examples of otherpre-formed toxins. A toxin may also be made within the body, often bymicrobes, but can be made by the patient's own cells, as occurs in viralinfection. Examples include toxins TcdA and TcdB, made by Clostridiumdificile bacterium and released into the intestinal lumen that killscells in the gastrointestinal tract, leading to severe, potentiallylife-threatening diarrhea. Another example is Shiga-like toxin orverotoxin that is produced by certain strains of Escherichia coli thatare frequently transmitted by contaminated undercooked meat, vegetables,or fruit and cause food-borne illness. Verotoxin is produced by thebacterium in the intestinal lumen, but then is absorbed into thebloodstream, where it is taken up by vascular endothelial cells, killingthem. Verotoxin preferentially targets the glomerulus, causing renalfailure and can lead to fatal hemolytic uremic syndrome. Yet anotherexample is the NSP4 toxin, produced by a patient's own cells followinggastrointestinal infection with the virus, rotavirus. Rotavirus is themost common cause of severe diarrhea in infants, killing more than600,000 patients a year. NSP4 acts as an enterotoxin, activating ionchannels in the colonic epithelium, causing a profuse secretorydiarrhea, without causing any structural damage. An excess of a normallynon-toxic substance, can also become a toxin when present at highconcentrations. An example includes cytokines, which are normallyproduced proteins of the immune system that at low levels are importantfor immune system function, but at high levels become inflammatorytoxins that cause cell death, severe inflammation, and organdysfunction. Toxins can also be formed by the metabolism of a non-toxicprecursor or protoxin. One example is amyloid precursor protein, anon-toxic protein produced in the body that when cleaved by enzymescalled secretases, can form the fragment beta-amyloid protein, a toxinthat can lead to the formation of amyloid plaques, the pathogenic rootcause of Alzheimer's disease. Non-toxic subunits or precursors can alsobecome toxins when combined with each other. An example of this is theproduction of lethal factor, edema factor, and protective antigen byBacillus anthracia, which is responsible for the disease anthrax. Bythemselves, they are non-toxic, but when combined together and takeninto the cell, these subunits form the deadly toxins, lethal toxin andedema toxin, causing cell death and cell lysis. Another example isStaphylococcus aureus alpha toxin, which is not toxic as a monomer, butbecomes a potent hemolytic toxin when seven monomers assemble into apore forming complex. A substance may also have no toxic effects when inone location in the body, but may become a toxin when placed in anotherpart of the body. An example of this is the gram negative bacterialendotoxin, lipopolysaccharide, which has little toxicity in theintestinal lumen, but causes septic shock if it escapes into thebloodstream. Toxins can also be common enzymes, such as lipases,amylases, trypsin, hyaluronidase, collagenase and others that cause cellor tissue damage when not appropriately regulated, or when active in aregion of the body that is not protected against their enzymatic action.When these substances help the spread or virulence of pathogens in thebody, they are classified as “virulence factors”. These are examples andnot meant to be limiting.

The term “gastrointestinal lumen” or “lumen” refers to the space orcavity within a gastrointestinal tract.

The term “gastrointestinal disorder” as used herein includes gastritis,Ménétrier disease, gastrointestinal ulceration, gastroenteritis,gastrointestinal inflammatory disease, enteric infection, gut-mucosalinjury, inflammatory bowel disease, celiac disease, and the like.

As used herein, the term “sorbent” includes adsorbents and absorbents.

As used herein, the term “physiologic fluids” are liquids that originatefrom the body and can include, but are not limited to, nasopharyngeal,oral, esophageal, gastric, pancreatic, hepatic, pleural, pericardial,peritoneal, intestinal, prostatic, seminal, vaginal secretions, as wellas tears, saliva, lung or bronchial secretions, mucus, bile, blood,lymph, plasma, serum, synovial fluid, cerebrospinal fluid, urine, andinterstitial, intracellular, and extracellular fluid, such as fluid thatexudes from burns or wounds.

As used herein, “carrier fluids” are exogenously administered liquidsthat include, but are not limited to, liquids administered orally, via afeeding tube, peritoneally, or rectally such as an enema or colonicwash.

As used herein, the singular forms “a,” “an,” and “the” include theplural, and reference to a particular numerical value includes at leastthat particular value, unless the context clearly dictates otherwise.When a range of values is expressed, another embodiment includes fromthe one particular value and/or to the other particular value.Similarly, when values are expressed as approximations, by use of theantecedent “about,” it will be understood that the particular valueforms another embodiment. All ranges are inclusive and combinable.

Unless defined otherwise, all other technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art pertinent to the methods and compositions described. Asused herein, the following terms and phrases have the meanings ascribedto them unless specified otherwise.

In addition, various references are identified below and areincorporated by reference in their entirety.

The present invention is based in-part on the discovery that byengineering pore structure that is defined by the ratio of pore sizerange to pore volume, biocompatible sorbents compositions can bemanufactured that sorb toxins, toxin precursors, toxin subunits, orvirulence factors, collectively termed “toxins” of a broad range ofsizes irrespective of other physical characteristics. The biocompatiblesorbents can be used to inhibit or reduce contamination by one or moretoxins when introduced into a biological substance, generally mammalian,and particularly human. However, the biological substance can also be amanufactured substance that supports proliferation or maintenance of amicro-organism. The sorbent compositions will be useful in reducingtoxins while working in conjunction with anti-microbial agents that killor halt the growth of microbes that are either directly producing thetoxin or causing the toxin to be produced. The sorbent compositions willbe also useful in situations where other treatments may not beunavailable for a variety of reasons. For example, the sorbents can beused in cases where the toxin has not been definitively identified.Another instance where the present biocompatible sorbent and methods maybe useful is when a toxin is identifiable, but not yet sufficientlytyped or cultured for targeted treatments. Another instance is whereanti-microbials cannot be used, for fear of greater toxin release whenthe microbe dies that will worsen the patient's condition.

The present biocompatible sorbent compositions are comprised of aplurality of pores. The biocompatible sorbents are designed to adsorb abroad range of toxins from less than 1 kDa to 1,000 kDa. While notintending to be bound by theory, it is believed the sorbent acts bysequestering molecules of a predetermined molecular weight within thepores. The size of a molecule that can be adsorbed by the polymer willincrease as the pore size of the polymer increases. Conversely, as thepore size is increased beyond the optimum pore size for adsorption of agiven molecule, adsorption of said protein may or will decrease.

In one embodiment a porous polymer that absorbs small to midsize proteinmolecules equal to or less than 50,000 Daltons (50 kDa) and excludesabsorption of large blood proteins comprises the pore structure suchthat the total pore volume of pore size in the range of 50 Å to 40,000 Åare in the range of 0.5 cc/g to 5.0 cc/g dry sorbent. The sorbent has apore structure such that the total pore volume of pore size in the rangeof 50 Å to 40,000 Å is greater than 0.5 cc/g to 5.0 cc/g dry sorbent;wherein the ratio of pore volume between 50 Å to 40,000 Å (porediameter) to pore volume between 100 Å to 1,000 Å (pore diameter) of thesorbent is smaller than 3:1.

In another embodiment a porous polymer that optimally absorbs midsize tolarge size protein molecules of approximately 300,000 Daltons andexcludes or minimizes absorption of very large blood proteins comprisesthe pore structure such that the total pore volume of pore size in therange of 50 Å to 40,000 Å are in the range of 0.5 cc/g to 5.0 cc/g drysorbent. The sorbent has a pore structure such that the total porevolume of pore size in the range of 50 Å to 40,000 Å is greater than 0.5cc/g to 5.0 cc/g dry sorbent; wherein the ratio of pore volume between50 Å to 40,000 Å (pore diameter) to pore volume between 1,000 Å to10,000 Å (pore diameter) of the sorbent is smaller than 2:1.

In another embodiment a porous polymer that optimally absorbs very largesize protein molecules equal to or less than 1,000,000 Daltons andexcludes or minimizes absorption of very large blood proteins comprisesthe pore structure such that the total pore volume of pore size in therange of 50 Å to 40,000 Å are in the range of 0.5 cc/g to 5.0 cc/g drysorbent. The sorbent has a pore structure such that the total porevolume of pore size in the range of 50 Å to 40,000 Å is greater than 0.5cc/g to 5.0 cc/g dry sorbent; wherein the ratio of pore volume between50 Å to 40,000 Å (pore diameter) to pore volume between 10,000 Å to40,000 Å (pore diameter) of the sorbent is smaller than 3:1.

The biocompatible sorbent is introduced into a biological substance. Theterm biological substance includes substances found in vivo, generallyfrom a mammal such as dogs, cats, rabbits, cows, sheep, horses, pigs andgoats, preferably a human. The biological substances are, for example,cells, or physiologic fluids such as saliva, nasopharyngeal, blood,plasma, serum, gastrointestinal fluid, bile, cerebrospinal fluid,pericardial, vaginal fluid, seminal fluid, prostatic fluid, peritonealfluid, pleural fluid, urine, synovial fluid, interstitial fluid,intracellular fluid or cytoplasm, lymph, bronchial secretions, mucus, orvitreous or aqueous humor. The term also includes substances thatsupport proliferation or maintenance of eukaryotic or prokaryotic cells,viruses, fungi, or protozoa ex vivo, such as culture media, biologicalmatrices, eggs and the like. The biocompatible sorbent and biologicalsubstance can be introduced and admixed either in vivo or ex vivo.

Toxins are most commonly organic substances such as proteins, peptides,carbohydrates, lipids, nucleic acids, and combinations thereof (e.g.multimeric or multi-subunit proteins, glycoproteins, glycolipids,lipoproteins, etc.). Toxins can also include organic chemicals (e.g.alkaloids), as well as certain inorganic molecules (e.g. cyanide).

Toxins can be produced by a wide range of organisms. For example, toxinscan be produced by micro-organisms such as bacteria, viruses, fungi andparasites. Toxins can also be formed by macro-organisms such as insects,fish, crustaceans, shellfish, amphibians, reptiles, birds, and mammals.Toxins can also be produced by vegetative matter such as plants, algae,and phytoplankton. Viruses typically contain the genetic code, via DNAor RNA, or other nucleic acid sequences, to direct the cells they infectto manufacture viral proteins, including toxins. Many toxins can also befound environmentally, the product of natural processes. Toxins can beformed by the assembly and/or modification of subunits that may or maynot be toxic by themselves. Toxins may be non-toxic substances at lowconcentrations, but becomes toxic at higher concentrations. Some toxinsmay be location specific. Toxins can also be byproducts of metabolism ofnon-toxic precursors. Toxins can also be artificially synthesized andmanufactured (e.g. biowarfare applications, etc).

Exogenous toxins can be ingested, injected, inhaled, absorbed by theskin or mucosal surfaces, such as the buccal or oral mucosa, thesublingual mucosa, the nasal mucosa, the sinuses, the nasopharynx, theoropharynx, the respiratory tract, the gastrointestinal tract, theurogenital tract, and the eye mucosa. Endogenous toxins can be producedanywhere in the body, such as in the blood, the gastrointestinal tract,the respiratory tract, the urogenital tract, within tissues, and withinbody cavities. Often, endogenous toxins can act locally, by disruptinglocal biologic processes or causing local disease, or toxins can actsystemically, disrupting systemic biologic processes, causing systemicdisease or organ dysfunction. Non-toxic precursors can also be absorbedfrom the environment or produced in the body, and are then converted inthe body into toxins via metabolism or other modification.

Toxins can cause pathology via a number of different mechanisms. Somecause the direct killing of cells. For example, brown recluse spidervenom contains a number of toxins that make it both cytolytic andhemolytic. These toxins are enzymes such as hyaluronidase,deoxyribonuclease, ribonuclease, alkaline phosphatase, and lipase.Sphingomyelinase D is thought to be the protein component responsiblefor most of the tissue destruction and hemolysis caused by brown reclusespider envenomation. The intense inflammatory response mediated byarachidonic acid, prostaglandins, and chemotactic infiltration ofneutrophils is amplified further by an intrinsic vascular cascadeinvolving the mediator C-reactive protein and complement activation.These and other factors contribute to the local and systemic reactionsof necrotic arachnidism. Others act by causing a disruption in normalcellular physiology. For example, anthrax edema factor binds withprotective antigen, to cause direct cell death.

One of the most common sources of toxins are from enteric pathogens suchas Clostridium difficile, Enterohemorrhagic E. coli (EHEC)) such as E.coli OE157:H7 or E. coli O104:H4, Vibrio cholera, Shigella dysenteriae,and rotovirus. These toxins can damage or kill cells of thegastrointestinal mucosa, and can alter gastrointestinal homeostasis,often resulting in diarrhea and vomiting that can lead to dehydration,malabsorption, and potentially even death, particularly in the young,the elderly, and immunocompromised subjects. These toxins can result inmore serious complications such as colitis, bloody diarrhea,gastrointestinal bleeding, a decrease in immune system function, toxicmegacolon, intestinal perforation, shock, sepsis and death. A compromiseof the gastrointestinal mucosa caused by infection and toxins, canfurther lead to the translocation of bacteria and toxins, such asendotoxin, from the intestinal tract, into the blood or body, which cantrigger or exacerbate sepsis. According to the CDC, there were 221,226cases of cholera in 45 countries in 2009, causing approximately 5,000deaths. These were caused predominantly by toxigenic, or toxinproducing, strains of cholera serogroups O1 and O139, that producedlarge amounts of cholera toxin. Some toxins can be absorbed from thegastrointestinal tract, such as botulinum, or shiga-like toxin (STX-1),and distributed to different parts of the body via blood, or lymph,causing disease.

Enteric pathogens are commonly found in the environment and includebacteria, viruses, parasites and plants. Pathogenesis may be directlyrelated to the organism (e.g. Yersinia enterocolitica, Norwalk virus),related to a toxin produced by organism (e.g. Clostridium difficile,Enterotoxigenic E. coli (ETEC)), caused by changes in cellular functionresulting in release of pro-inflammatory cytokines (e.g.,Enteropathogenic E. coli (EPEC), Camplylobacter jejuni), or acombination of these of mechanisms (e.g. Vibrio cholera, Shigelladysenteriae).

Embodiments of the present invention can be used to inhibit or reducecontamination by toxins across a broad range of molecular weights and apre-formed toxin or a toxin formed in the presence of the biologicalsubstance. An example of a toxin formed in the presence of a biologicalsubstance is a bacterial endotoxin, such as lipopolysaccharide (LPS).The appearance of LPS in the host bloodstream is believed to lead to theendogenous production of a variety of host factors that directly andindirectly mediate the toxicity of LPS. These host-derived mediatorsinclude many now well-recognized inflammatory cytokines, endocrinehormones, and a number of other endogenous factors such as leukotrienesand platelet activating factor. Cytokines such as TNF-α, IL-1, and IFN-γare released from stimulated macrophages and T lymphocytes as a resultof infection by a variety of microorganisms, including bacteria,viruses, fungi, and parasites. The interacting factors comprise thecytokine cascade. TNF-alpha has been observed to stimulate production ofother types of cytokines. IL-1 induces responses observed ininflammation in general, such as fever, increase of leukocytes,activation of lymphocytes, and induction of biosynthesis of acute phaseprotein in liver. However, at high levels, these cytokines and otherscan be inflammatory toxins and cause undesirable effects such ascapillary leak, cell death via apoptosis, hemodynamic instability, organdysfunction, and cachexia.

As another example, in bacterial infections, cytokines such as IL-8 actas a signal that attracts white blood cells such as neutrophils to theregion of cytokine expression. In general, the release of enzymes andsuperoxide anions by neutrophils is essential for destroying theinfecting bacteria. However, if cytokine expression causes neutrophilsto invade, for example, the lungs, release of neutrophil enzymes andoxygen radicals can result in the development of adult respiratorydistress syndrome (ARDS), which can be lethal.

In certain embodiments, the toxin is from one or more diverse biologicalsources. Biological sources can comprise one or more bacteria, viruses,fungi, or parasites as shown in Table 1, a non-exclusive list of toxins,toxin subunits, and their representative pathogens.

TABLE 1 Toxins Exotoxin or Enterotoxin Genus Species Toxin MWEnterotoxigenic Enterotoxigenic 2 kDa and STa and STb E. coli 5.2 kDa,(heat stable (ETEC) respectively enterotoxin) StaphylococcalStaphylococcus S. aureus 23-29 kDa toxin B Alpha toxin Staphylococcus S.aureus 33 to 85 kDa Toxic shock Staphylococcus S. aureus 22 kDa syndrometoxin (TSST-1) Clostridium Clostridium C. perfringens 35 kDa perfringensenterotoxin C. perfringens Clostridium C. perfringens 43 kDa Alpha toxinAerolysin Aeromonas A. hydrophila 52 KDa Pseudomonas Pseudomonas P.aeruginosa 66 kDa Exotoxin A Shiga-like toxin EscherichiaEnterohemorrhagic 69 kDa (STX-1, STX-2; E. coli verotoxin) (EHEC) Shigatoxin Shigella dysenteria 70 kDa Cholera Toxin Vibrio Vibrio cholerae 84kDa Enterotoxigenic Escherichia Enterotoxigenic 86 kDa LT (heat labileE. ecoli enterotoxin) (ETEC) Lipopoly- Gram negative 10 kDa, upsaccharide bacteria to 1000 kDa Endotoxin aggregated Lipoteichoic acidGram positive 10 kDa Endotoxin bacteria Cyanotoxins Cyanobacteria Variedsizes Pertussis toxin Bordetella B. pertussis 105 kDa Tetanus ToxinClostridium C. tetani 150 kDa Botulinum toxin Clostridium C. botulium150 kDa C. diff toxin B Clostridium C. difficile 250-270 kDa (TcdB) C.diff toxin A Clostridium C. difficile 308 kDa (TcdA)

Table 2 is a non-exclusive list of viral toxins where the presentsorbent can be used to reduce or inhibit contamination of a biologicalsubstance.

TABLE 2 VIRUS Toxin Family Virus MW Rotavirus NSP4 toxin ReoviridaeRotavirus 28 kDa

Table 3 is a non-exclusive list of fungal toxins where use of thepresent sorbent can reduce or inhibit contamination of a biologicalsubstance.

TABLE 3 FUNGUS Toxin Genus Species MW adhesins Candida C. albicans, C.esophagitis Aflatoxin Aspergillus A. flavus 312-346 Daltons and other A.parasiticus mycotoxins Amatoxin Amanita A. phalloides,     900 DaltonsA. brunnescens

Table 4 is a non-exclusive list of plant-derived toxins where use of thepresent sorbent can reduce or inhibit contamination of a biologicalsubstance.

TABLE 4 PLANT Toxin Common Name Genus and Species MW Ricin toxin CastorBean Ricinis communis 65 kDa Pyrrolizidine Legumes, fescue Alkaloidsgrass, contaminated milk and flour Phytohaemagglutinin Kidney BeansPhaseolus vulgaris Vomitoxin Grains

Table 5 is a non-exclusive list of animal-derived toxins where use ofthe present sorbent can reduce or inhibit contamination of a biologicalsubstance.

TABLE 5 ANIMAL DERIVED Toxin Example of Source MW Ciguatera fishGrouper, red snapper, toxin amberjack, barracuda Tetrodotoxin Pufferfish Scombroid (Histamine) Fish: bluefin, tuna, skipjack, marlin,mackerel Shellfish toxin Mussels, scallops, shrimps, oysters (shellfish)Cytokines (TNF-α, Subject's cells, egg cells 10 kDa, up IL-1, IL-6, to1000 kDa IL-8, IL-10) aggregated Hemoglobin

Furthermore, the compositions of biocompatible sorbents can comprise amixture of sorbents with different pore structures. Such a mixture willbe advantageous, for example, when the pathogen has not beendefinitively identified or there is reason to suspect there are multiplepathogens associated with the gastroenteritis, such as with theingestion of contaminated water.

In another example, the present methods and sorbents can be used intreating infections with Clostridium difficile via oral or rectaladministration. C. difficile is commonly acquired in hospitals worldwideand other long-term healthcare settings. It is a leading cause of severediarrhea in these institutions. In the hospital setting the consequencesinclude, longer stays, more readmissions and higher costs. (Glouliouriset al., Clin. Med., 11:75-79, 2011). C. difficile spores are excreted inlarge numbers by patients and contamination can survive for months oryears. The bacteria are generally inhibited by the normal flora of thegastrointestinal system, but an imbalance or disruption in the colonicflora, allows for C. difficile to proliferate and produce toxinsresulting in diarrhea, and possibly fever and colitis. The availabletreatment is metronidazole and vancomycin, but reoccurrence is commonbecause full recovery requires re-establishing the normal intestinalflora. C. difficile produce two toxins, Toxin A and Toxin B, both ofwhich are pathogenic. (Lyerly et al., Clin. Microbiol. Rev., 1(1):1-18,1988) The proteins are quite large, both Toxins A and B are greater than250,000 under reduced conditions and have been described ranging from400,000 to 600,000 Daltons for Toxin A (Krivan and Wilkins, Infect.Immun., 55:1873-1877, 1987; Bano et al., Rev. Infect. Dis. 6:S11-S201984; Bano et al., Biochem. Int. 2:629-635, 1981), and 360,000 to500,000 Daltons for Toxin B (Lyerly et al., Infect. Immun. 54:70-76,1986).

In another example, the present methods and sorbents can be used intreating bacterial toxins in the blood using a hemocompatible sorbent inan extracorporeal hemoperfusion system. Standard hemodialysis,hemofiltration and charcoal hemoperfusion are not capable of removingtoxins larger than approximately 10 kDa. High molecular weight cutofffilters, and mid-molecular weight sorbent cartridges are not ideallysuited for broad, particularly large (>60 kDa) toxin removal. Thissorbent system, in conjunction with antibiotics, could be used to removepathogen derived toxins from blood, plasma or scrum in a range of lessthan 1 kDa to more than 400 kDa. Examples of infections that alsoproduce blood-borne toxins include Staphylococcus aureus andmethicillin-resistant Staphlococus aureus (MRSA) that produce alphatoxin or toxic shock syndrome toxin-1; enterohemorrhagic E. coli enteralinfection associated with systemic shiga-like toxin toxemia; Clostridiumperfringens infection with alpha toxin production causing necrotizingfasciitis; Aeromonas wound infection with aerolysin toxin production,and others.

In some embodiments, the sorbent comprises a coated polymer comprisingat least one crosslinking agent and at least one dispersing agent.

Some preferred coated polymers comprise at least one crosslinking agentand at least one dispersing agent. Suitable dispersing agents includehydroxyethyl cellulose, hydroxypopyl cellulose, poly(hydroxyethylmethacrylate), poly(hydroxyethyl acrylate), poly(hydroxypropylmethacrylate), poly(hydroxypropyl acrylate), poly(dimethylaminoethylmethacrylate), poly(dimethylaminoethyl acrylate), poly(diethylamimoethylmethacrylate), poly(diethylaminoethyl acrylate), poly(vinyl alcohol),poly(N-vinylpyrrolidinone), salts of poly(methacrylic acid), and saltsof poly(acrylic acid) and mixtures thereof.

Suitable crosslinking agents include divinylbenzene, trivinylbenzene,divinylnaphthalene, trivinylcyclohexane, divinylsulfone,trimethylolpropane trimethacrylate, trimethylolpropane dimethacrylate,trimethylolpropane triacrylate, trimethylolpropane diacrylate,pentaerythrital dimethacrylates, pentaerythrital trimethacrylates,pentaerythrital, tetramethacrylates, pentaerythritol diacrylates,pentaerythritol triiacrylates, pentaerythritol tetraacrylates,dipentaerythritol dimethacrylates, dipentaerythritol trimethacrylates,dipentaerythritol tetramethacrylates, dipentaerythritol diacrylates,dipentaerythritol triacrylates, dipentaerythritol tetraacrylates,divinylformamide and mixtures thereof. Preferably, the polymer isdeveloped simultaneously with the formation of the coating, such thatthe dispersing agent gets chemically bound to the surface of thepolymer.

The use of an organic solvent as a porogen or pore-former, and theresulting phase separation induced during polymerization yield porouspolymers. Some preferred porogens are Benzyl alcohol, Cyclohexane,Cyclohexanol, Cyclohexanol/toluene mixtures, Cyclohexanone, Decane,Decane/toluene mixtures, Di-2-ethylhexylphosphoric acid, Di-2-ethylhexylphthalate, 2-Ethyl-1-hexanoic acid, 2-Ethyl-1-hexanol,2-Ethyl-1-hexanolin-heptane mixtures, 2-Ethyl-1-hexanol/toluenemixtures, Isoamyl alcohol, n-Heptane, n-Heptane/ethylacetate,n-Heptane/isoamyl acetate, n-Heptane/tetraline mixtures,n-Heptane/toluene mixtures, n-Hexane/toluene mixtures, Pentanol,Poly(styrene-co-methyl methacrylate)/dibutyl phthalate,Polystyrene/2-ethyl-1-hexanol mixtures, Polystyrene/dibutyl phthalate,Polystyrene/n-hexane mixtures, Polystyrene/toluene mixtures, Toluene,Tri-n-butylphosphate, 1,2,3-Trichloropropane/2-ethyl-1-hexanol mixtures,2,2,4-Trimethyl pentane (isooctane), Trimethyl pentane/toluene mixtures,Poly(propylene glycol)/toluene mixtures Poly(propyleneglycol)/cyclohexanol mixtures, and Poly(propyleneglycol)/2-Ethyl-1-hexanol mixtures

Preferred sorbents comprise polymers derived from one or more monomersselected from divinylbenzene and ethylvinylbezene, styrene,ethylstyrene, acrylonitrile, butyl methacrylate, octyl methacrylate,butyl acrylate, octyl acrylate, cetyl methacrylate, cetyl acrylate,ethyl methacrylate, ethyl acrylate, vinyltoluene, vinylnaphthalene,vinylbenzyl alcohol, vinylformamide, methyl methacrylate, methylacrylate, trivinylbenzene, divinylnaphthalene, trivinylcyclohexane,divinylsulfone, trimethylolpropane trimethacrylate, trimethylolpropanedimethacrylate, trimethylolpropane triacrylate, trimethylolpropanediacrylate, pentaerythritol dimethacrylate, pentaerythritoltrimethacrylate, pentaerythritol tetramethacrylate, pentaerythritoldiacrylate, pentaerythritol triiacrylate, pentaerythritol tetraacrylate,dipentaerythritol dimethacrylate, dipentaerythritol trimethacrylate,dipentaerythritol tetramethacrylate, dipentaerythritol diacrylate,dipentaerythritol triacrylate, dipentaerythritol tetraacrylate,divinylformamide and mixtures thereof.

Some preferred polymers comprise ion exchange polymers.

Some preferred polymers comprise cellulosic polymers. Suitable polymersinclude cross-linked dextran gels such as Sephadex™.

Certain preferred polymers comprise porous highly crosslinked styrene ordivinylbenzene copolymers. Some of these copolymers comprise amacroporous or mesoporous styrene-divinylbenzene-ethylstyrene copolymersubjected to a partial chloromethylation to a chlorine content of up to7% molecular weight. Other of these polymers are a hypercrosslinkedpolystyrene produced from crosslinked styrene copolymers by an extensivechloromethylation and a subsequent post-crosslinking by treating with aFriedel-Crafts catalyst in a swollen state. Yet other of these polymersare a hypercrosslinked polystyrene produced from crosslinked styrenecopolymers by an extensive additional post-crosslinking in a swollenstate with bifunctional crosslinking agents selected from the groupcomprising of monochlorodimethyl ether and p-xylilene dichloride

Some polymers useful in the practice of the invention are hydrophilicself-wetting polymers that can be administered as dry powder containinghydrophilic functional groups such as, amines, hydroxyl, sulfonate, andcarboxyl groups.

Certain polymers useful in the invention are macroporous polymersprepared from the polymerizable monomers of styrene, divinylbenzene,ethylvinylbenzene, and the acrylate and methacrylate monomers such asthose listed below by manufacturer. Rohm and Haas Company, (now part ofDow Chemical Company): (i) macroporous polymeric sorbents such asAmberlite™ XAD-1, Amberlite™ XAD-2, Amberlite™ XAD-4, Amberlite™ XAD-7,Amberlite™ XAD-7HP, Amberlite™ XAD-8, Amberlite™ XAD-16, Amberlite™XAD-16 HP, Amberlite™ XAD-18, Amberlite™ XAD-200, Amberlite™ XAD-1180,Amberlite™ XAD-2000, Amberlite™ XAD-2005, Amberlite™ XAD-2010,Amberlite™ XAD-761, and Amberlite™ m XE-305, and chromatographic gradesorbents such as Amberchrom™ m CG 71,s,m,c, Amberchrom™ CG 161,s,m,c,Amberchrom™ CG 300,s,m,c, and Amberchrom™ CG 1000,s,m,c. Dow ChemicalCompany: Dowex™ Optipore™ L-493, Dowex™ Optipore™ V-493, Dowex™Optipore™ V-502, Dowex™ Optipore™ L-285, Dowex™ Optipore™ L-323, andDowex™ m Optipore™ m V-503. Lanxess (formerly Bayer and Sybron):Lewatit™ VPOC 1064 MD PH, Lewatit™ VPOC 1163, Lewatit™ OC EP 63,Lewatit™ S 6328A, Lewatit™ OC 1066, and Lewatit™ 60/150 MIBK. MitsubishiChemical Corporation: Diaion™ HP 10, Diaion™ HP 20, Diaion™ HP 21,Diaion™ HP 30, Diaion™ HP 40, Diaion™ HP 50, Diaion™ SP70, Diaion™ SP205, Diaion™ SP 206, Diaion™ SP 207, Diaion™ SP 700, Diaion™ SP 800,Diaion™ SP 825, Diaion™ SP 850, Diaion™ SP 875, Diaion™ HP 1MG, Diaion™HP 2MG, Diaion™ CHP 55A, Diaion™ CHP 55Y, Diaion™ CHP 20A, Diaion™ CHP20Y, Diaion™ CHP 2MGY, Diaion™ CHP 20P, Diaion™ HP 20SS, Diaion™ SP20SS, and Diaion™ SP 207SS. Purolite Company: Purosorb™ AP 250 andPurosorb™ AP 400.

The present invention does not rely on charge or a ligand-receptorcomplex binding reaction to inhibit or reduce pathogen toxicity. Apolymer using acid functional group(s) attached to the polymer backboneto bind Clostridium difficile Toxin A and Toxin B is described by BaconKurtz et al. (U.S. Pat. No. 6,890,523). The interaction in Kurtz isionic where a hydrophobic or hydrophilic group attached to the polymerbinds the toxin. Chamot el al. (US Patent Application 2006/009169)describe using inorganic polymer particles linked to a toxin bindingmoiety comprised of oligosaccharide sequences that bind C. difficileToxin A and Toxin B. Also described is a toxin binding surface pore size2× larger than toxin diameter. Chamot described oligosaccharide moietiesthat bind toxins to form a ligand/receptor-like complex.

The polymer materials used as the sorbent are generally notmetabolizable by human and animal, but may be synthesized from materialscharacterized as being a biodegradable polymer, a resorbable polymer, orboth. Certain polymers may be irregular or regular shaped particulatessuch as powders, beads, or other forms with a diameter in the range of0.1 micron meters to 2 centimeters.

The polymers used in the instant invention preferably have abiocompatible and hemocompatible exterior surface coatings but are notabsolutely necessary, especially in certain circumstances, such as oralor rectal administration. Certain of these coatings are covalently boundto the polymer particle (beads, for example) by free-radical grafting.The free-radical grafting may occur, for example, during thetransformation of the monomer droplets into polymer beads. Thedispersant coating and stabilizing the monomer droplets becomescovalently bound to the droplet surface as the monomers within thedroplets polymerize and are converted into polymer. Biocompatible andhemocompatible exterior surface coatings can be covalently grafted ontothe preformed polymer beads if the dispersant used in the suspensionpolymerization is not one that imparts biocompatibility orhemocompatibility. Grafting of biocompatible and hemocompatible coatingsonto preformed polymer beads is carried out by activating free-radicalinitiators in the presence of either the monomers or low molecularweight oligomers of the polymers that impart biocompatibility orhemocompatibility to the surface coating.

The route of administration can be systemic or localized. In certainembodiments, the compositions may be given orally, rectally or via afeeding tube. The sorbent can be supplied as a dry powder or other dryparticulate capable of being wetted externally or internally in thealimentary canal, including in the gastric or enteric environment, withor without the addition of wetting agents such as ethyl or isopropylalcohol, potable liquids such as water, or other carrier fluid. Otherpossible routes of administration include subcutaneous or transdermaldelivery. In some embodiments, administration is topical. Such methodsinclude ophthalmic administration, administration to skin or wounds,direct administration into a body cavity or joint, and delivery tomucous membranes such as nasal, oral, vaginal and rectal delivery orother delivery to the alimentary canal. In some embodiments, thetreatment is extracorporeal. Extracorporeal administration would includeremoval of inflammatory mediators from blood or physiologic fluids bycirculating the fluids through a device containing sorbent and returningit back to the body. In some embodiments, such methods include local orsystemic administration through a parenteral route. Parenteraladministration includes intravenous, intraarterial, subcutaneous,intraperitoneal or intramuscular injection or infusion; or intracranial(including intrathecal or intraventricular, administration).

The sorbent may be formulated as for example, a powder, a tablet, acapsule, a solution, a slurry, an emulsion, a suppository, or in a foodsubstance. The sorbent may be packaged in portable bottles, vials,blister packs, bags, pouches, or other container that allows for eithersingle or multiple dosages. Depending on the use, the sorbent may besterile or non-sterile. The polymer may be sterilized by standardmethods. Such methods are well known to those skilled in the art. Thetherapeutically effective amount can be administered in a series ofdoses separated by appropriate time intervals, such as hours. Thecompositions of the instant invention may be administered by methodswell known to those skilled in the art.

The foregoing description is provided for the purpose of explanation andis not to be construed as limiting the invention. While the inventionmay have been described with reference to preferred embodiments orpreferred methods, it is understood that the words which have been usedherein are words of description and illustration, rather than words oflimitation. Furthermore, although the invention has been describedherein with reference to particular materials structure, methods,compositions and embodiments, the invention is not intended to belimited to the particulars disclosed herein, as the invention extends toall structures, methods, compositions and uses that are within the scopeof the appended claims. Further, several advantages have been describedthat flow from the composition and methods; the present invention is notlimited to composition and methods that encompass any or all of theseadvantages. Those skilled in biocompatible polymer technology, havingthe benefit of the teachings of this specification, may effect numerousmodifications to the invention as described herein, and changes can bemade without departing from the scope and spirit of the invention asdefined by the appended claims. Furthermore, any features of onedescribed embodiment can be applicable to the other embodimentsdescribed herein. For example, any features or advantages related to thedesign of the biocompatible polymers with respect to discussion of aparticular toxin absorption embodiment can be applicable to any of theother toxin absorption embodiments described herein.

The invention is further illustrated by the following non-limitingexamples.

EXAMPLES Examples 1-18

Eighteen porous polymeric adsorbents are characterized for their porestructures and their syntheses are described in Examples 1-18. The porestructure characterization is given in Example 19.

The synthesis process consists of (1) preparing the aqueous phase, (2)preparing the organic phase, (3) carrying out the suspensionpolymerization, (4) purifying the resulting porous polymeric adsorbentproduct (work-up), and (5) addition of a hemo-compatible coating.

The following synthesis procedure is generalized to fit all sampleswhich were made. The synthetic process varied between each polymersample; refer to Table 6, following the generalized procedure, in orderto see specific run conditions for each example.

Reactor Setup. A 5 L or 0.5 L kettle reactor was fitted with anover-head stirrer, a water cooled condenser, a multi-level stirrerblade, a thermocouple, and a bubbler. For the 0.5 L kettles, a gasketwas installed between the top lid and bottom kettle. The 5 L set-ups hada baffle plate assembly and two flat rubber gaskets installed betweenthe top lid and bottom kettle. All unused ports were capped with theappropriate plug. Temperature was controlled with a heating mantle whichwas regulated by a temperature controller fitted with theabove-mentioned thermocouple.

Polymerization. Polyvinyl alcohol (“PVA”) was dispersed in one half ofthe water charge at room temperature (RT) and then heated to 70° C. Theremaining salts: MSP, DSP, TSP, & Sodium Nitrite were then dissolved inthe remainder of the water charge. The PVA solution and salts solutionwere each added to the reactor and heated to the desired reactiontemperature with stirring. The pre-mixed organic phase, including theinitiator, was poured into the reactor onto the aqueous phase with thestirring speed set at the revolutions per minute (“rpm”) for formationof appropriate droplet size. Once the temperature reached the set-point,the reaction timer was set for 16 hours and started and the reaction wasallowed to proceed.

Work-up. Solvent level marked. After cooling, the solvent was siphonedout to the bead level. The beads were then washed 5 times with 50°C.-70° C. water at a rate of 1 bed volume per half hour. The followingsteps in the work-up were skipped if the polymer was modified (See Table6). The beads were then washed 3 times with RT methanol at a rate of 1bed volume per 10 minutes. The polymer was extracted via a soxhletapparatus overnight. The polymer was steam stripped for 8 hours. Afterthe steam strip was completed, the polymer was rewet in isopropylalcohol and then sieved with purified water to the desired particlesize. The polymer was then dried in an oven at 100° C.

Modification Setup. A kettle reactor was fitted with an over-headstirrer, a multi-level stirrer blade, and a thermocouple. All unusedports were capped with the appropriate plug and one open hose adapter asa vent. A gasket was installed between the top lid and bottom kettle.Temperature was controlled with a heating mantle regulated by atemperature controlled fitted with the above-mentioned thermocouple.

Modification Reaction. Polymer was washed 10 times with isopropylalcohol at approximately 1 bed volume per hour and then 10 times withpurified water at approximately 1 bed volume per hour. The polymer wassieved to the desired particle size and added to the reactor setup.Excess water was siphoned to just above bed level and the charged waterwas then added. The temperature controller was set to 40° C. and thenstarted. The overhead stirrer was started as well. Each reagent wasadded while the system was ramping up to the 40° C. set point. AmmoniumPersulfate (AMPS) in water was added when the temperature was between30° C. to 34° C. NNNN-Tetramethylethylenediamine (TMED) and water wereadded between 35° C. and 36° C. Vinylpyrrolidinone (VP) and water wereadded between 39° C. and 40° C. The two hour reaction timer was startedwhen the temperature reached 40° C.; the reaction was allowed toproceed. After cooling, the solvent was siphoned out to the bead level.The beads were then washed 3 times with RT water at a rate of 1 bedvolume per half hour. The beads were steam stripped for 6 hours. Thebeads were rewet in isopropyl alcohol and washed ten times in purifiedH₂O. The polymer was then dried in an oven at 100° C.

This process resulted in a clean, dry adsorbent in the form ofspherical, porous polymer beads.

TABLE 6 Synthesis Conditions for Examples 1-18. Example Example ExampleExample Example 1 2 3 4 5 TDG- TDG- RJR- TDG- RJR- 057-64 071-167090-030 057-118 090-013 Run Conditions Kettle Size 0.5 5.0 0.5 0.5 0.5Reaction Temperature 80 80 80 87 87 Aqueous Phase Charges Item Charge, gCharge, g Charge, g Charge, g Charge, g Ultrapure Water 231.26 1734.47231.26 231.26 231.26 Polyvinyl Alcohol (PVA) 0.68 5.06 0.68 0.68 0.68Monosodium Phosphate (MSP) 0.71 5.34 0.71 0.71 0.71 Disodium Phosphate(DSP) 2.36 17.71 2.36 2.36 2.36 Trisodium Phosphate (TSP) 1.47 10.991.47 1.47 1.47 Sodium Nitrite 0.01 0.05 0.01 0.01 0.01 Total 236.491773.62 236.49 236.49 236.49 Organic Phase Charges Item Charge, gCharge, g Charge, g Charge, g Charge, g Divinylbenzene (DVB)(63%) 129.55592.92 83.03 83.03 83.03 Toluene 0.00 390.48 0.00 0.00 0.00 Isooctane0.00 448.47 0.00 0.00 0.00 Cyclohexanol 102.82 0.00 143.45 151.00 151.00PPG 0.00 0.00 7.55 0.00 0.00 Benzoyl Peroxide (BPO)(97%) 1.32 4.49 0.840.84 0.84 Total, w/o BPO 232.37 1431.87 234.03 234.03 234.03 Work-UpMethanol Washes 3 N/A 3 3 3 Soxhlet Solvent Acetone N/A Acetone AcetoneAcetone Sieve Size (μm) 300-600 300-600 300-600 300-600 300-600Modification Amount of Polymer being modified, N/A 500 N/A N/A 400 mLCharged Water, mL N/A 180 N/A N/A 144 Ammonium Persulfate (AMPS), g N/A3.2 N/A N/A 2.7 in Water for Addition, mL N/A 28 N/A N/A 22NNNN-Tetramethylethylenediamine N/A 3.4 N/A N/A 2.9 (TMED), g in Waterfor Addition, mL N/A 14 N/A N/A 11 Vinylpyrrolidinone (VP), g N/A 1.7N/A N/A 1.4 in Water for Addition, mL N/A 42 N/A N/A 33 Example ExampleExample Example Example 6 7 8 9 10 RJR- RJR- RJR- RJR- RJR- 090-014090-016 090-023 090-087 090-091 Run Conditions Kettle Size 5.0 5.0 0.50.5 0.5 Reaction Temperature 80 87 80 80 80 Aqueous Phase Charges ItemCharge, g Charge, g Charge, g Charge, g Charge, g Ultrapure Water1500.00 1734.47 231.26 231.26 231.26 Polyvinyl Alcohol (PVA) 4.38 5.060.68 0.68 0.68 Monosodium Phosphate (MSP) 4.63 5.34 0.71 0.71 0.71Disodium Phosphate (DSP) 15.31 17.71 2.36 2.36 2.36 Trisodium Phosphate(TSP) 9.50 10.99 1.47 1.47 1.47 Sodium Nitrite 0.05 0.05 0.01 0.01 0.01Total 1533.87 1773.62 236.49 236.49 236.49 Organic Phase Charges ItemCharge, g Charge, g Charge, g Charge, g Charge, g Divinylbenzene(DVB)(63%) 1373.00 581.23 129.55 94.73 106.38 Toluene 0.00 0.00 0.000.00 0.00 Isooctane 0.00 0.00 0.00 0.00 0.00 Cyclohexanol 0.00 1057.0177.11 128.47 114.14 PPG 0.00 0.00 25.70 10.42 12.68 Benzoyl Peroxide(BPO)(97%) 6.91 5.91 1.32 0.96 1.08 Total, w/o BPO 1373.00 1638.24232.36 233.62 233.20 Work-Up Methanol Washes N/A 3 3 3 3 Soxhlet SolventN/A Acetone Acetone Acetone Acetone Sieve Size (μm) 106-212 106-212300-600 300-600 300-600 Modification Amount of Polymer being modified,500 600 N/A N/A N/A mL Charged Water, mL 180 216 N/A N/A N/A AmmoniumPersulfate (AMPS), g 0.0324 4.1 N/A N/A N/A in Water for Addition, mL 2833 N/A N/A N/A NNNN-Tetramethylethylenediamine 0.0169 4.3 N/A N/A N/A(TMED), g in Water for Addition, mL 42 17 N/A N/A N/A Vinylpyrrolidinone(VP), g 0.0344 2.1 N/A N/A N/A in Water for Addition, mL 14 50 N/A N/AN/A Example Example Example Example 11 12 13 14 RJR- RJR- RT- TDG-090-136 090-137 075-14-1 057-145 Run Conditions Kettle Size 0.5 0.5 5.05.0 Reaction Temperature 80 80 80 80 Aqueous Phase Charges Item Charge,g Charge, g Charge, g Charge, g Ultrapure Water 231.26 231.26 1500.001734.47 Polyvinyl Alcohol (PVA) 0.68 0.68 4.38 5.06 Monosodium Phosphate(MSP) 0.71 0.71 4.63 5.34 Disodium Phosphate (DSP) 2.36 2.36 15.31 17.71Trisodium Phosphate (TSP) 1.47 1.47 9.50 10.99 Sodium Nitrite 0.01 0.010.05 0.05 Total 236.49 236.49 1533.87 1773.62 Organic Phase Charges ItemCharge, g Charge, g Charge, g Charge, g Divinylbenzene (DVB)(63%) 106.3894.73 1373.00 592.92 Toluene 0.00 0.00 0.00 390.48 Isooctane 0.00 0.000.00 448.47 Cyclohexanol 115.73 130.21 0.00 0.00 PPG 11.10 8.68 0.000.00 Benzoyl Peroxide (BPO)(97%) 1.08 0.96 6.91 4.49 Total, w/o BPO233.21 233.62 1373.00 1431.87 Work-Up Methanol Washes 3 3 N/A N/ASoxhlet Solvent Acetone Acetone N/A N/A Sieve Size (μm) 300-600 300-600300-600 45-106 Modification Amount of Polymer being modified, N/A N/AN/A 2000 mL Charged Water, mL N/A N/A N/A 721 Ammonium Persulfate(AMPS), g N/A N/A N/A 12.9 in Water for Addition, mL N/A N/A N/A 111NNNN-Tetramethylethylenediamine N/A N/A N/A 13.8 (TMED), g in Water forAddition, mL N/A N/A N/A 55 Vinylpyrrolidinone (VP), g N/A N/A N/A 6.7in Water for Addition, mL N/A N/A N/A 166 Example Example ExampleExample 15 16 17 18 RJR- RJR- RJR- RJR- 090-178 090-021 090-187 090-031Run Conditions Kettle Size 0.5 0.5 0.5 0.5 Reaction Temperature 80 80 8080 Aqueous Phase Charges Item Charge, g Charge, g Charge, g Charge, gUltrapure Water 231.26 231.26 231.26 231.26 Polyvinyl Alcohol (PVA) 0.680.68 0.68 0.68 Monosodium Phosphate (MSP) 0.71 0.71 0.71 0.71 DisodiumPhosphate (DSP) 2.36 2.36 2.36 2.36 Trisodium Phosphate (TSP) 1.47 1.471.47 1.47 Sodium Nitrite 0.01 0.01 0.01 0.01 Total 236.49 236.49 236.49236.49 Organic Phase Charges Item Charge, g Charge, g Charge, g Charge,g Divinylbenzene (DVB)(63%) 106.38 83.03 106.38 106.38 Toluene 0.00 0.000.00 0.00 Isooctane 0.00 0.00 0.00 0.00 Cyclohexanol 109.07 113.25104.63 107.80 PPG 17.76 37.75 22.19 19.02 Benzoyl Peroxide (BPO)(97%)1.08 0.84 1.08 1.08 Total, w/o BPO 233.21 234.03 233.20 233.20 Work-UpMethanol Washes 3 3 3 3 Soxhlet Solvent Acetone Acetone Acetone AcetoneSieve Size (μm) 300-600 300-600 300-600 300-600 Modification Amount ofPolymer being modified, N/A N/A N/A N/A mL Charged Water, mL N/A N/A N/AN/A Ammonium Persulfate (AMPS), g N/A N/A N/A N/A in Water for Addition,mL N/A N/A N/A N/A NNNN-Tetramethylethylenediamine N/A N/A N/A N/A(TMED), g in Water for Addition, mL N/A N/A N/A N/A Vinylpyrrolidinone(VP), g N/A N/A N/A N/A in Water for Addition, mL N/A N/A N/A N/A

Example 19 Pore Structure Characterization

The pore structures of the adsorbent polymers were analyzed with eithera Micromeritics AutoPore IV 9500 V1.09 a Mercury Penetrometer (HgIntrusion instrument) or a Micromeritics ASAP 2010 instrument (N₂Desorption). The results are shown in FIG. 1 where the pore volume isplotted as a function of the pore diameter. FIG. 1 is the logdifferential pore structures for Examples 1-18.

Example 20 Pore Structure Characterization

The pore volume is divided up into categories within pore size rangesfor each of the sorbent polymers and these values are provided in Tables7 and 8. In the first range, the capacity pore volume is that porevolume that is accessible to protein sorption and consists of the porevolume in pores larger than 100 Å in diameter. The effective pore volumeis that pore volume that is selectively accessible to proteins smallerthan approximately 50,000 Daltons and consists of pore diameters withinthe range of 100 to 1000 Å in diameter. The oversized pore volume is thepore volume accessible to proteins larger than approximately 50,000Daltons and consists of the pore volume in pores larger than 1000 Å indiameter. The undersize pore volume is the pore volume in pores smallerthan 100 Å diameter and is not accessible to proteins larger than about10,000 Daltons.

In the second range, the capacity pore volume is that pore volume thatis accessible to protein sorption and consists of the pore volume inpores larger than 1,000 Å in diameter. The effective pore volume is thatpore volume that is selectively accessible to proteins smaller thanabout 300,000 Daltons and consists of pore diameters within the range of1000 to 10000 Å in diameter. The oversized pore volume is the porevolume accessible to proteins larger than 300,000 Daltons and consistsof the pore volume in pores larger than 1000 Å in diameter. Theundersize pore volume is the pore volume in pores smaller than 1,000 Ådiameter and is not accessible to proteins larger than about 10,000Daltons.

In the third range, the capacity pore volume is that pore volume that isaccessible to protein sorption and consists of the pore volume in poreslarger than 500 Å in diameter. The effective pore volume is that porevolume that is selectively accessible to proteins smaller than 1,000,000Daltons and consists of pore diameters within the range of 10,000 to40,000 Å in diameter. The oversized pore volume is the pore volumeaccessible to proteins larger than 1,000,000 Daltons and consists of thepore volume in pores larger than 40,000 Å in diameter. The undersizepore volume is the pore volume in pores smaller than 10,000 Å diameterand is not accessible to proteins larger than about 40,000 Daltons.

Table 7 provides the pore volumes and pore volume ratios for Examples1-18.

TABLE 7 Pore Volume Pore Volume Pore Volume Pore Volume (cc/g) of pores,(cc/g) of pores, (cc/g) of pores, (cc/g) of pores, diameter rangediameter range diameter range diameter range Polymer Name 50-40000 Å100-1000 Å 1000-10000 Å 10000-40000 Å TDG-057-064 0.82 0.71 0.00 0.00TDG-071-167 1.75 0.80 0.78 0.05 RJR-090-030 2.25 0.32 1.72 0.17TDG-057-118 2.54 0.88 1.26 0.32 RJR-090-013 2.54 0.88 1.26 0.32RJR-090-014 N/A N/A N/A N/A RJR-090-016 2.27 0.91 1.08 0.20 RJR-090-0230.95 0.10 0.81 0.02 RJR-090-087 1.69 0.20 1.38 0.06 RJR-090-091 1.460.25 1.20 0.01 RJR-090-136 1.68 0.41 1.18 0.07 RJR-090-137 1.91 0.241.53 0.10 RT-075-14-1 N/A N/A N/A N/A TDG-057-145 1.75 0.80 0.78 0.05RJR-090-178 1.36 0.15 1.12 0.05 RJR-090-021 1.52 0.04 0.21 1.24RJR-090-187 0.88 0.05 0.13 0.68 RJR-090-031 1.22 0.09 0.52 0.57

Table 8 provides the pore volume ratios for Examples 1-18.

TABLE 8 Ratio of Pore Ratio of Pore Ratio of Pore Volume Volume VolumeBetween Between Between 50-4000 Å 50-40000 Å 50-40000 Å 0 to pore volumeto pore volume to pore volume between between between Polymer Name100-1000 Å 1000-10000 Å 10000-40000 Å TDG-057-064 1.2:1 219.8:1  N/ATDG-071-167 2.2:1 2.2:1  2.2:1 RJR-090-030 6.9:1 1.3:1 13.4:1TDG-057-118 2.9:1 2.0:1  8.0:1 RJR-090-013 2.9:1 2.0:1  8.0:1RJR-090-014 N/A N/A N/A RJR-090-016 2.5:1 2.1:1 11.4:1 RJR-090-023 9.1:11.2:1 52.9:1 RJR-090-087 8.4:1 1.2:1 27.2:1 RJR-090-091 6.0:1 1.2:1163.1:1  RJR-090-136 4.1:1 1.4:1 25.7:1 RJR-090-137 7.9:1 1.3:1 19.2:1RT-075-14-1 N/A N/A N/A TDG-057-145 2.2:1 2.2:1 37.6:1 RJR-090-178 9.0:11.2:1 25.6:1 RJR-090-021 42.1:1  7.1:1  1.2:1 RJR-090-187 19.4:1  6.7:1 1.3:1 RJR-090-031 13.9:1  2.3:1  2.1:1

Example 21 In Vitro C. difficile Toxin A(rTcdA)

The main objective of this study was to evaluate the ability of polymerbeads (Porous beads ID's: TDG-057-118, RJR-090-136, RJR-090-091,RJR-090-137, RJR-090-023 and RJR-090-178, and non-porous bead ID:RT-075-1-14) to bind Clostridium difficile rTcdA. Eight types of beads,those with and without pores, were utilized. rTcdA was evaluated at aconcentration of 100 μg/ml. No beads, and 20 μL of each non-porous andbeads with pores were incubated with 100 (ideally 64.65) μg/ml of rTcdAat a 0.3 ml final working volume containing phosphate buffer saline in a2-mL screw cap tube. Immediately after addition of toxins, one tube ineach group containing no beads, beads without pores or porous beadsstood for 0.583 h, allowing the beads to settle. A 225 μl sample wastaken from these tubes. These were designated as the 0.583 h samples.Tubes from which the 1.5 and 2.5 h samples were taken were placed on atube roller. A 225 μl sample was removed from these tubes. All sampleswere stored at −20° C. until use. Following collection of all samples,the protein concentration remaining in each sample was evaluated usingthe BCA (bicinchoninic acid) protein assay (Thermo Scientific, Cat. NO.23225). Results are shown below. Beads TDG 057-118 demonstrated the besttoxin removal, as well as RJR-090-136.

Table 9 provides the weight of polymers used for Example 21.

TABLE 9 Amount Bulk Wet Bead Weight Sample Name (μL) Density (mg/μL)(mg) RJR-090-023 20 0.2794 5.6 RJR-090-091 20 0.3863 7.7 RJR-090-136 200.3240 6.5 RJR-090-137 20 0.2794 5.6 RJR-090-178 20 0.3487 7.0TDG-057-118 20 0.2239 4.5 RT-075-14-1 20 0.8037 16.1

The C. difficile Toxin A adsorption results are shown in table 10

TABLE 10 Expected Concentration (μg/mL) at Designated Time Point (h)Conc. BCA % Toxin Removal (μg/mL) Sample 0.00 h 0.58 h 1.50 h 2.50 h 0 h0.583 h 1.5 h 2.5 h 100 No Bead 64.65 83.46 70.53 73.29 0% 0%  0%  0%RT-075-14-1 64.65 70.54 61.14 69.76 0% 0%  5%  0% TDG-057-118 64.6563.14 20.55 14.05 0% 2% 68% 78% RJR-090-136 64.65 62.89 18.54 16.36 0%3% 71% 75% RJR-090-091 64.65 67.21 31.72 16.41 0% 0% 51% 75% RJR-090-13764.65 74.87 32.68 16.68 0% 0% 49% 74% RJR-090-023 64.65 67.49 27.03 18.70% 0% 58% 71% RJR-090-178 64.65 70.06 21.88 15.63 0% 0% 66% 76%

FIG. 2 shows C. difficile Toxin A removal over time.

Example 22 In Vitro C. difficile Toxin B (rTcdB)

The main objective of this study was to evaluate the ability of polymerbeads (Porous bead ID: RJR-090-016, and non-porous bead ID: RJR-090-014)to bind and remove Clostridium difficile rTcdB toxin in vitro at toxinconcentrations of 25 and 100 μg/mL, as compared to a no-bead control. Inbrief, either no beads, or a fixed volume (46 μL each) of eithernon-porous (≈37.0 μg dry bead weight) or porous beads (≈12.1 μg dry beadweight) were incubated with either 25 or 100 μg/ml of rTcdB in phosphatebuffered saline at a 0.3 ml final working volume in a 2-mL screw capmicrofuge tube. The experiment was performed to keep the interstitialvolume (outside of the beads) constant at 0.3 mL. The weight of theporous beads reflects their high degree of porosity compared to thenon-porous beads Immediately after addition of toxin, one tube in eachgroup containing either no beads, non-porous bead or porous beads wasallowed to sit without agitation for 45 minutes to allow the beads tosettle by gravity. A 225 μl sample was taken from these tubes and storedat −20° C. These were designated as the 0.75 h samples. Sample tubes,designated 1.75 h and 2.75 h respectively, were mixed continuously on atube roller. At time-points of 1.0 and 2.0 hours after the start of theexperiment, respectively, tubes were removed from the roller and set ina rack for 45 min for the beads to settle. A 225 μl sample was removedfrom these tubes. All samples were stored at −20° C. until use.Following collection of all samples, the protein concentration remainingin each sample was evaluated using the BCA (bicinchoninic acid) proteinassay (Thermo Scientific, Cat. NO. 23225). The results shown below inTable 11 represent the concentration of rTcdB toxin in each of thealiquoted samples, over time. Although non-porous beads demonstrate somebinding to the toxin, the contribution of the porous bead is clear,particularly at higher concentrations, where saturation of thenon-porous beads is evident, but is not observed in the porous beads.

The C. difficile Toxin B adsorption results are shown in Table 11.

TABLE 11 Starting Concentration (μg/mL) at Concentration Designated TimePoint (h) % Removal (μg/mL) Sample 0.75 1.75 2.75 (at T_(2.75 h)) 25 NoBead 25.2 25.5 32.7 ~0% Non-Porous 13.7 5.3 7.8 69% Bead Porous Bead16.3 Not Detected 0.8 97% 100 No Bead 110.4 128.3 105.9  0% Non-Porous118.5 92.5 94.2  6% Bead Porous Bead 90.9 8.4 2.3 98%

Example 23 In Vitro C. difficile Toxin B (rTcdB)

The main objective of this study was to evaluate the ability ofCytosorbents' Beads (Porous beads ID's: TDG-057-118, RJR-090-136,RJR-090-091, RJR-090-137, RJR-090-023 and RJR-090-087, and non-porousbead ID: RT-075-1-14) to bind Clostridium difficile rTcdB. Seven typesof beads, those with and without pores, and a no bead control wereutilized. rTcdB was evaluated at a concentration of 100 μg/ml. No beads,and 20 μL (dry weight of sample, see table 11 below) of each non-porousand beads with pores were incubated with 100 (ideally 99.76) μg/ml ofrTcdB at a 0.3 ml final working volume containing phosphate buffersaline in a 2-mL screw cap tube.

The weight of polymers used for Example 19 is shown in Table 12.

TABLE 12 Amount Bulk Wet Bead Weight Sample Name (μL) Density (mg/μL)(mg) RJR-090-023 20 0.2794 5.6 RJR-090-087 20 0.3487 7.0 RJR-090-091 200.3863 7.7 RJR-090-136 20 0.3240 6.5 RJR-090-137 20 0.2794 5.6TDG-057-118 20 0.2239 4.5 RT-075-14-1 20 0.8037 16.1

Immediately after addition of toxins, one tube in each group containingno beads, bead without pores or porous beads were stood for 0.583 h,allowing the beads to settle. A 225 μl sample was taken from thesetubes. These were designated as the 0.583 h samples. Tubes from whichthe 1.5 and 2.5 h samples were taken were placed on a tube roller. A 225μl sample was removed from these tubes. All samples were stored at −20°C. until use. Following collection of all samples, the proteinconcentration remaining in each sample was evaluated using the BCA(bicinchoninic acid) protein assay (Thermo Scientific, Cat. NO. 23225).Results are shown below. Beads RJR-090-136 demonstrated the best toxinremoval, followed by TDG 057-118 and/or RJR 090-137.

The C. difficile Toxin B adsorption results are shown in Table 13.

TABLE 13 C. diff toxin B Concentration (μg/mL) at Expected DesignatedTime Point (h) Conc. BCA % of Toxin Removal (μg/mL) Sample 0 h .583 h1.5 h 2.5 h 0 h .583 h 1.5 h 2.5 h 100 No Bead 99.76 83.53 81.94 71.960% 16% 18% 28% RT-075-14-1 99.76 122.14 81.58 93.07 0%  0% 18%  7%TDG-057-118 99.76 91.18 39.28 21.05 0%  9% 61% 79% RJR-090-136 99.7682.83 36.43 17.81 0% 17% 63% 82% RJR-090-091 99.76 93.73 62.79 30.58 0% 6% 37% 69% RJR-090-137 99.76 85.28 52.45 19 0% 15% 47% 81% RJR-090-02399.76 71.52 61.08 38.05 0% 28% 39% 62% RJR-090-087 99.76 92.92 59.1523.11 0%  7% 41% 77%

FIG. 3 shows C. difficile Toxin B removal over time.

Example 24 In Vitro Botulinum Neurotoxin Type A1 (BoNT/A1) Study

The main objective of this study was to evaluate the ability of polymerbeads (Porous bead ID: TDG-057-118, and non-porous bead ID:RT-075-14-1). Two types of beads, those with and without pores, wereutilized. BoNT/A1 was evaluated at concentrations of 10, 50 and 100μg/ml in phosphate buffered saline. No beads, or a fixed volume of 40 μLeach of either non-porous (≈32.1 μg dry bead weight) or porous beads(≈5.5 μg dry bead weight) were incubated with either 10, 50 or 100 μg/mlof BoNT/A1 at a 0.3 ml final working volume in a 2-mL screw capmicrofuge tube. The experiment was performed to keep the interstitialvolume (outside of the beads) constant at 0.3 mL. The weight of theporous beads reflects their high degree of porosity compared to thenon-porous beads. Immediately after addition of BoNT/A1, a 225 μl samplewas taken from one tube in each group containing no beads, non-porousbeads, or porous beads and stored at −20° C. These were designated asthe 0 h samples. Tubes correlating to the 1 and 2 h samples were placedon a tube roller and mixed continuously. After incubation at roomtemperature of either 1 or 2 hours, a 225 μl sample was removed from theappropriate tubes. All samples were stored at −20° C. until use.Following collection of all samples, the protein concentration remainingin each sample was evaluated using the BCA (bicinchoninic acid) proteinassay (Thermo Scientific, Cat. NO. 23225). As can be seen in the resultslisted in Table 14, at concentrations up to 100 μg/mL, the porous beadsremoved more than 95% of the BoNT/A1 toxin, compared to either theno-bead control or the non-porous beads.

The Botulinum Neurotoxin Type A1 adsorption results are shown in Table14.

TABLE 14 Starting Concentration (μg/mL) at % Concentration DesignatedTime Point (h) Removal (μg/mL) Sample 0 h 1 h 2 h (at T_(2 h)) 10 NoBead 10.2 6.9 7.2 29% Non-Porous 10.2 7.4 5.6 45% Bead Porous Bead 7.2Not Not 100%  Detected Detected 50 No Bead 43.4 47.2 42.8  1% Non-Porous47.3 43.7 38.6 18% Bead Porous Bead 37.9 1.7 1.5 96% 100 No Bead 117.696.8 105.8 10% Non-Porous 94.1 120.3 105.8  0% Bead Porous Bead 74.6 4.53.5 95%

Example 25 In Vitro Shiga Like Toxin 1 Study

The main objective of this study was to evaluate the ability ofCytoSorbents polymer beads (Porous bead ID: TDG-071-167, Large Pore BeadID: RJR-090-013 and non-porous bead ID: RT-075-14-1) to bind Shiga LikeToxin 1. Three types of beads, those with and without pores, wereutilized. Shiga Like Toxin 1 was evaluated at concentrations of 50 and100 μg/ml in phosphate buffered saline. No beads, and 42 μL of porous(≈12.5 μg dry bead weight), 42 μL of large pore (≈9.5 μg dry beadweight) and 42 μL of non-porous beads (≈33.8 μg dry bead weight) wereincubated with either 50 or 100 μg/ml of Shiga Like Toxin 1 at a 0.3 mlfinal working volume in a 2-mL screw cap microfuge tube.

Immediately after addition of Shiga Like Toxin 1, a 225 μl sample wastaken from one tube in each groups containing no beads, non-porousbeads, or porous beads and stored at −20° C. These were designated asthe 0.25 h samples. Tubes correlating to the 1.25 and 2.25 h sampleswere placed on a tube roller and mixed continuously. After incubation atroom temperature of either 1.25 or 2.25 hours, a 225 μl sample wasremoved from the appropriate tubes. All samples were stored at −20° C.until use. Following collection of all samples, the proteinconcentration remaining in each sample was evaluated using the BCA(bicinchoninic acid) protein assay (Thermo Scientific, Cat. NO. 23225).As can be seen, the both standard and large porous beads have a betterkinetics of removal in contrast to the non-porous beads.

The Shiga Like Toxin 1 adsorption results are shown in Table 15.

TABLE 15 Stx1 Starting Concentration (μg/ml) at Concentration DesignatedTime Point % removal % removal (μg/ml) Sample 0.25 h 1.25 h 2.25 h atT0.25 h at T2.25 h 50 No Bead Control 36.4 23.46 27.77 27% 44%RT-075-14-1 23.38 24.71 21.52 53% 57% TDG-071-167 20.93 Not 0.99 58% 98%Determined RJR-090-013 20.93 Not 0.52 58% 99% Determined 100 No BeadControl 53.58 42.23 47.78 46% 52% RT-075-14-1 41.51 37.6  49.65 58% 50%TDG-071-167 39.55 3.7 0.8 60% 99% RJR-090-013 38.74 Not 0.71 61% 99%Determined

Example 26 In Vitro Shiga Like Toxin 2 Study

The main objective of this study was to evaluate the ability ofCytoSorbents polymer beads (Porous bead ID: TDG-071-167, Large Pore BeadID: RJR-090-013 and non-porous bead ID: RT-075-14-1) to bind Shiga LikeToxin 2. Three types of beads, those with and without pores, wereutilized. Shiga Like Toxin 2 was evaluated at concentrations of 50 and100 μg/ml in phosphate buffered saline. No beads, and 42 μL of porous(≈12.5 μg dry bead weight), 42 μL of large pore (≈9.5 μg dry beadweight) and 42 μL of non-porous beads (≈33.8 μg dry bead weight) wereincubated with either 50 or 100 μg/ml of Shiga Like Toxin 2 at a 0.3 mlfinal working volume in a 2-mL screw cap microfuge tube Immediatelyafter addition of Shiga Like Toxin 2, a 225 μl sample was taken from onetube in each groups containing no beads, non-porous beads, or porousbeads and stored at −20° C. These were designated as the 0.25 h samples.Tubes correlating to the 1.25 and 2.25 h samples were placed on a tuberoller and mixed continuously. After incubation at room temperature ofeither 1.25 or 2.25 hours, a 225 μl sample was removed from theappropriate tubes. All samples were stored at −20° C. until use.Following collection of all samples, the protein concentration remainingin each sample was evaluated using the BCA (bicinchoninic acid) proteinassay. As can be seen, the both standard and large porous beads have abetter kinetics of removal in contrast to the non-porous beads.

The Shiga Like Toxin 2 adsorption results are shown in Table 16.

TABLE 16 Stx2 Starting Concentration (μg/ml) at Concentration DesignatedTime Point % removal % removal (μg/ml) Sample 0.25 h 1.25 h 2.25 h atT0.25 h at T2.25 h 50 No Bead Control 35.22 35.63 32.86 30% 34%RT-075-14-1 32 30.57 31.32 36% 37% TDG-071-167 22.45 0.54 1.9 55% 96%RJR-090-013 27.75 1.03 0.95 45% 98% 100 No Bead Control 69.4 70.02 69.0331% 31% RT-075-14-1 60.51 65.58 55.75 39% 44% TDG-071-167 59.24 4.33 2.141% 98% RJR-090-013 54.8 2.38 1.06 45% 99%

Example 27 In Vitro Ricin Toxin Study

The main objective of this study was to evaluate the ability ofCytoSorbents polymer beads (Small Porous bead ID: TDG-057-145, Modified,Batch 1, −106/+45, Large porous bead ID: RJR-090-016 and non-porous beadID: RJR-090-014) to bind ricin toxin. Three types of beads, those withand without pores, were utilized. Ricin toxin was evaluated atconcentrations of 100 and 1000 μg/ml in phosphate buffered saline. Nobeads, and 43 μL of porous beads (≈14.6 μg dry bead weight), 43 μL oflarge pore (≈11.3 μg dry bead weight), and 44 μL of non-porous beads(≈35.4 μg dry bead weight) were incubated with either 100 or 1000 μg/mlof ricin toxin at a 0.3 ml final working volume in a 2-mL screw capmicrofuge tube. Immediately after addition of ricin toxin, a 225 μlsample was taken from one tube in each groups containing no beads,non-porous beads, or porous beads and stored at −20° C. These weredesignated as the 0.75 h samples. Tubes correlating to the 1.75 and 2.75h samples were placed on a tube roller and mixed continuously. Afterincubation at room temperature of either 1.75 or 2.75 hours, a 225 μlsample was removed from the appropriate tubes. All samples were storedat −20° C. until use. Following collection of all samples, the proteinconcentration remaining in each sample was evaluated using the BCA(bicinchoninic acid) protein assay (Thermo Scientific, Cat. NO. 23225).As can be seen, the small porous beads have a better kinetics of removalin contrast to the large porous beads initially. No-bead control or thenon-porous beads removed no toxins and no more than 9%, respectively.

The Ricin Toxin adsorption results are shown in Table 17.

TABLE 17 Ricin Toxin Starting Concentration (μg/ml) at ConcentrationDesignated Time Point % removal % removal (μg/ml) Sample 0.75 h 1.75 h2.75 h at T0.75 h at T2.75 h 100 No Bead Control 111.21 114.2 117.69  0% 0% RJR-090-014 90.8 104.36 109.02  9%  0% TDG-057-145 2.67 2.05 2.3897% 98% RJR-090-016 12.77 1.28 0.62 87% 99% 1000 No Bead Control 1136.331246.77 1025.9  0%  0% RJR-090-014 1240.62 1217.81 1341.14  0%  0%TDG-057-145 170.4 8.7 10.49 83% 99% RJR-090-016 353.44 11.98 9.18 65%99%

Example 28 In Vitro Cholera Toxin Study

The main objective of this study was to evaluate the ability ofCytoSorbents™ polymer beads (Small Porous bead ID: TDG-057-145,Modified, Batch 1, −106/+45, Large porous beads bead ID: RJR-090-016 andnon-porous bead ID: RJR-090-014) to bind cholera toxin. Three types ofbeads, those with and without pores, were utilized. Cholera toxin wasevaluated at concentrations of 50 and 100 μg/ml in phosphate bufferedsaline. No beads, and 43 μL of standard porous beads (≈14.6 μg dry beadweight), 43 μL of large pore (≈11.3 μg dry bead weight), and 44 μL ofnon-porous beads (≈35.4 μg dry bead weight) were incubated with either50 or 100 μg/ml of cholera toxin at a 0.3 ml final working volume in a2-mL screw cap micro fuge tube. Immediately after addition of choleratoxin, a 225 μl sample was taken from one tube in each groups containingno beads, non-porous beads, or porous beads and stored at −20° C. Thesewere designated as the 0.75 h samples. Tubes correlating to the 1.75 and2.75 h samples were placed on a tube roller and mixed continuously.After incubation at room temperature of either 1.75 or 2.75 hours, a 225μl sample was removed from the appropriate tubes. All samples werestored at −20° C. until use. Following collection of all samples, theprotein concentration remaining in each sample was evaluated using theBCA (bicinchoninic acid) protein assay (Thermo Scientific, Cat. NO.23225). As can be seen, the small porous beads have a better kinetics ofremoval in contrast to the large porous beads. No-bead control or thenon-porous beads removed no toxins and less than 25%, respectively.

The cholera toxin adsorption results are shown Table 18.

TABLE 18 Starting Concentration (μg/mL) at Concentration Designated TimePoint (h) % Removal % Removal (μg/mL) Sample 0.75 1.75 2.75 (atT_(0.75 h)) (at T_(2.75 h)) 50 No Bead Control 63.82 57.29 65.16  0%  0%RJR-090-014 38.54 37.24 38.55 23% 23% TDG-057-145 4.37 3.24 2.41 91% 95%RJR-090-016 6.38 3.24 3.41 87% 93% 100 No Bead Control 96.06 128.22113.45  4%  0% RJR-090-014 76.13 91.23 79.42 24% 21% TDG-057-145 5.944.91 3.24 94% 97% RJR-090-016 10.64 4.1 2.35 89% 98%

Example 29 In Vitro C. perfringens Enterotoxin Study

The main objective of this study was to evaluate the ability ofCytoSorbents polymer beads (Porous bead ID: TDG-071-167, Large Pore BeadID: TDG-057-118 and non-porous bead ID: RT-075-14-1) to bind C.perfringens enterotoxin. Three types of beads, those with and withoutpores, were utilized. C. perfringens enterotoxin was evaluated atconcentrations of 50 and 100 (ideally 11.46 and 31.42) μg/ml inphosphate buffered saline. No beads, and 40 μL of porous beads (≈11.9 μgdry bead weight), 40 μL of large pore (≈9.0 μg dry bead weight) and 40μL of non-porous beads (≈32.1 μg dry bead weight) were incubated witheither 50 or 100 (ideally 11.46 and 31.42) μg/ml of C. perfringensenterotoxin at a 0.3 ml final working volume in a 2-mL screw capmicrofuge tube. Immediately after addition of C. perfringensenterotoxin, a 225 μl sample was taken from one tube in each groupscontaining no beads, non-porous beads, or porous beads and stored at−20° C. These were designated as the 0.5 h samples. Tubes correlating tothe 1.5 and 2.5 h samples were placed on a tube roller and mixedcontinuously. After incubation at room temperature of either 1.5 or 2.5hours, a 225 μl sample was removed from the appropriate tubes. Allsamples were stored at −20° C. until use. Following collection of allsamples, the protein concentration remaining in each sample wasevaluated using the BCA (bicinchoninic acid) protein assay (ThermoScientific, Cat. NO. 23225). As can be seen, the large porous beads havea better kinetics of removal from 31.42 mg/mL of toxin in contrast tothe standard and non-porous beads. No-bead control or the non-porousbeads did not remove any toxin.

The C. perfringens Enterotoxin adsorption results are shown in Table 19.

TABLE 19 C. perfringens Enterotoxin Concentration (μg/ml) at ExpectedDesignated Time Point Concentration BCA % removal % removal (μg/ml)Sample 0 h 0.5 h 1.5 h 2.5 h at T0.5 h at T2.5 h 50 No Bead 11.46 18.0513.94 15.43  0%  0% RT-075-14-1 — 12.4 11.82 17.59  0%  0% TDG-071-167 —4.22 0.35 2.63 63% 77% TDG-057-118 — 7.81 0.98 1.74 32% 85% 100 No Bead31.42 34.91 34.73 34.78  0%  0% RT-075-14-1 — 36.25 35.82 37.31  0%  0%TDG-071-167 — 27.73 0.54 2.82 12% 91% TDG-057-118 — 19.52 0.52 2.25 38%93%

Example 30 In Vitro Staphylococcus Enterotoxin B Study

The main objective of this study was to evaluate the ability ofCytoSorbents polymer beads (Small Porous bead ID: TDG-057-145, andnon-porous bead ID: RJR-090-014) to bind Staphylococcus enterotoxin B.Two types of beads, those with and without pores, were utilized.Staphylococcus enterotoxin B was evaluated at concentrations of 50 and100 (ideally 43.02 and 97.85) μg/ml in phosphate buffered saline. Nobeads, and 43 μL of porous (≈14.6 μg dry weight) and 44 μL of non-porousbeads (≈35.4 μg dry weight) were incubated with either 50 or 100(ideally 43.02 and 97.85)μg/ml of Staphylococcus enterotoxin at a 0.3 mlfinal working volume in a 2-mL screw cap microfuge tube. Immediatelyafter addition of Staphylococcus enterotoxin, a 225 μL sample was takenfrom one tube in each groups containing no beads, non-porous beads, orporous beads and stored at −20° C. These were designated as the 0.75 hsamples. Tubes correlating to the 1.75 and 2.75 h samples were placed ona tube roller and mixed continuously. After incubation at roomtemperature of either 1.75 or 2.75 hours, a 225 μL sample was removedfrom the appropriate tubes. All samples were stored at −20° C. untiluse. Following collection of all samples, the protein concentrationremaining in each sample was evaluated using the BCA (bicinchoninicacid) protein assay. As can be seen, the small porous beads have abetter kinetics of removal (greater than or equal to 98% by 0.75 h) incontrast to the non-porous beads. No-bead control or the non-porousbeads did not remove toxins as efficiently.

The Staphylococcus Enterotoxin B adsorption results are shown in Table20.

TABLE 20 Staph Enterotoxin Starting Concentration (μg/ml) atConcentration Designated Time Point % removal % removal (μg/ml) Sample0.75 h 1.75 h 2.75 h at T0.75 h at T2.75 h 50 No Bead Control 43.0259.09 42.13 14% 16% RJR-090-014 31 38.04 23.38 38% 53% TDG-057-145 1.241.36 ND 98% 100%  100 No Bead Control 97.85 100.3 95.43  2%  5%RJR-090-014 54.08 72.36 70.8 46% 29% TDG-057-145 1.77 1.99 0.92 98% 99%

Example 31 In Vitro Staphylococcus aureus α-Hemolysin Example 31: InVitro Staphylococcus aureus α-Hemolysin

The main objective of this study was to evaluate the ability of twodifferent CytoSorbents porous bead types (Small Pore: RJR-100-144; LargePore: RJR-100-168) and non-porous beads (RJR-090-158) to bindStaphylococcus aureus α-Hemolysin. S. aureus α-Hemolysin was evaluatedat concentrations of 50 and 100 μg/ml in phosphate buffered saline. Nobeads, and 40 μL of porous beads (≈11.9 μg dry bead weight), 40 μL oflarge pore (≈9.0 μg dry bead weight), and 40 μL of non-porous beads(≈32.1 μg dry bead weight) were incubated with either 50 or 100 μg/ml oftoxin at a 0.3 ml final working volume in a 1.5-mL screw cap tube.Immediately after addition of Staphylococcus aureus α-Hemolysinenterotoxin, a 225 μl sample was taken from one tube in each groupcontaining no beads, non-porous beads, or porous beads and immediatelystored at −20° C. Tubes correlating to the 0.5 h, 1.5 h, and 2.5 hourssamples were placed on a tube roller and mixed continuously. Afterincubation at room temperature of either 0.5 h, 1.5 h, or 2.5 hours, a225 μl sample was removed from the appropriate tubes. All samples werestored at −20° C. until use. Following collection of all samples, theprotein concentration remaining in each sample was evaluated using theBCA (bicinchoninic acid) protein assay (Thermo Scientific, Cat. NO.23225). The BCA assay indicates that both porous polymers have betterkinetics of removal than the no bead control and the non-porous beads.

S. aureus α-Hemolysin adsorption results via BCA protein assay andNanoDrop are shown in Table 21 below.

TABLE 21 Staphylococcus aureus α- Hemolysin Concentration (μg/mL) atExpected Designated Time Point Concentration BCA % removal % removal(μg/ml) Sample 0 h 0.5 h 1.5 h 2.5 h at T0.5 h at T2.5 h 50 No Bead(PBS) 57.31 53.93 54.66 48.63  6% 15% RJR-090-158 47.14 54.66 49.5 18%14% RJR-100-144 23.8 4.15 0 58% 100%  RJR-100-168 42.47 0 0 26% 100% 100 No Bead (PBS) 119.96 113.44 91.62 108.89  5%  9% RJR-090-158 97.14103.45 95.8 19% 20% RJR-100-144 39.45 16.44 3.28 67% 97% RJR-100-16866.36 0 0 45% 100% 

Example 32 In Vitro Escherichia Coli STa Toxin

The main objective of this study was to evaluate the ability of variousCytoSorbents porous bead types (bead #1: SFA-102-106, bead #2: CytoSorbLot 08311, bead #3: TDG-057-118) and non-porous beads (RT-075-14-1) tobind Escherichia coli STa toxin. Escherichia coli STa toxin wasevaluated at concentrations of 50 and 100 μg/mL in phosphate bufferedsaline. No beads, 40 μL of SFA-102-106 (≈9.0 μg dry bead weight), 40 μLof CytoSorb Lot 083111 (≈11.9 μg dry bead weight), 40 μL of TDG-057-118(≈9.0 μg dry bead weight), and 40 μL of non-porous beads (≈32.1 μg drybead weight) were incubated with either 50 or 100 μg/ml of Escherichiacoli STa toxin at a 0.3 ml final working volume in a 1.5-mL screw captube. Immediately after addition of Escherichia coli STa toxin, a 225 μlsample was taken from one tube in each groups containing no beads,non-porous beads, or porous beads and immediately stored at −20° C.Tubes correlating to the 0.5 h, 1.5 h, and 2.5 hours samples were placedon a tube roller and mixed continuously. After incubation at roomtemperature of either 0.5 h, 1.5 h, or 2.5 hours, a 225 μl sample wasremoved from the appropriate tubes. All samples were stored at −20° C.until use. Following collection of all samples, the proteinconcentration remaining in each sample was evaluated using the BCA(bicinchoninic acid) protein assay (Thermo Scientific, Cat. NO. 23225)The BCA assay indicates that all three porous polymers have betterkinetics of removal than the no bead control and the non-porous beads.

The Escherichia coli STa toxin adsorption results are shown in Table 22.

TABLE 22 Concentration (μg/ml) at Expected Designated Time PointConcentration BCA % removal % removal (μg/mL) Sample 0 h 0.5 h 1.5 h 2.5h at T0.5 h at T2.5 h 50 No Bead 71.72 58.24 55.54 55.55 19% 23%RT-075-14-1 — 50.16 54.85 71.72 30%  0% SFA-102-106 — 55.55 7.03 4.3423% 94% Standard-083111 — 58.24 8.73 1.64 19% 98% TDG-057-118 — 55.554.33 1.05 58% 99% 100 No Bead 133.71  128.32 125.62 136.41  4%  0%RT-075-14-1 — 131.01 128.32 133.71  2%  0% SFA-102-106 — 106.75 9.724.34 20% 97% Standard-083111 — 106.8 20.51 12.42 20% 91% TDG-057-118 —112.14 70.03 4.44 16% 97%

What is claimed:
 1. A method of reducing contamination by one or moretoxins in a biological substance, said method comprising: a. contactingthe biological substance with an effective amount of a sorbent capableof sorbing the toxin, wherein the sorbent comprises a plurality of poresranging from 50 Å to 40,000 Å with a pore volume of 0.5 cc/g to 5.0 cc/gand a size of 0.05 mm to 2 cm, wherein the sorbent comprises a coatedpolymer comprising at least one crosslinking agent and wherein saidtoxins comprise endogenous toxins having a molecular weight of greaterthan 50,000 Daltons; and b. sorbing the toxin; wherein the sorbent has:(i) a ratio of pore volume between 50 Å to 40,000 Å (pore diameter) topore volume between 100 Å to 1,000 Å (pore diameter) of the sorbent issmaller than 3:1; or (ii) a ratio of pore volume between 50 Å to 40,000Å (pore diameter) to pore volume between 1,000 Å to 10,000 Å (porediameter) of the sorbent is smaller than 2:1; or (iii) a ratio of porevolume between 50 Å to 40,000 Å (pore diameter) to pore volume between10,000 Å to 40,000 Å (pore diameter) of the sorbent is smaller than 3:1;and wherein the toxin is an exotoxin produced by a micro-organism thatcan comprise one or more bacteria, viruses, fungi or parasites; whereinsaid toxins comprise proteins, peptides, carbohydrates, lipids, nucleicacids, or combinations thereof.
 2. The method of claim 1, wherein thesorbent is biocompatible.
 3. The method of claim 1, wherein the polymeris a microporous polymeric sorbent.
 4. The method of claim 1, whereinthe sorbing occurs in vivo.
 5. The method of claim 1, wherein thesorbing occurs ex vivo.
 6. The method of claim 1, wherein the biologicalsubstance comprises cells or physiologic fluids such as saliva,nasopharyngeal fluid, blood, plasma, serum, saliva, gastrointestinalfluid, bile, cerebrospinal fluid, pericardial, vaginal fluid, seminalfluid, prostatic fluid, peritoneal fluid, pleural fluid, urine, synovialfluid, interstitial fluid, intracellular fluid, extracellular fluid,lymph, mucus, or vitreous humor.
 7. The method of claim 1, wherein themethod further comprises the steps of producing or purifying a bloodproduct or biologic.
 8. The method of claim 7, wherein the blood productcomprises whole blood, packed red blood cells, platelets, plasma,cryoprecipitate, white blood cells, pluripotent stem cells, T-cells,B-cells, or other cells of myloid or lymphoid origin and theirprogenitors.
 9. The method of claim 1, wherein contamination by thetoxin is systemic or localized.
 10. The method of claim 1, wherein thesorbent is introduced through a body cavity.
 11. The method of claim 10,wherein the sorbent is introduced orally, vaginally, rectally ornasally, through a feeding tube or topically.
 12. The method of claim 1,wherein the sorbent is introduced by hemoperfusion.
 13. The method ofclaim 1, wherein the sorbent is used for extracorporeal treatment of abiological substance comprising saliva, blood, plasma, serum,gastrointestinal fluid, cerebrospinal fluid, vaginal fluid, peritonealfluid, pleural fluid, urine, synovial fluid, lymph, alveolar mucus orvitreous humor.
 14. The method of claim 1, wherein the sorbent has apore structure such that the total pore volume of pore size in the rangeof 50 Å to 40,000 Å is greater than 0.5 cc/g to 5.0 cc/g dry sorbent;wherein the ratio of pore volume between 50 Å to 40,000 Å (porediameter) to pore volume between 100 Å to 1,000 Å (pore diameter) of thesorbent is smaller than 3.1.
 15. The method of claim 1, wherein thesorbent has a pore structure such that the total pore volume of poresize in the range of 50 Å to 40,000 Å is greater than 0.5 cc/g to 5.0cc/g dry sorbent; wherein the ratio of pore volume between 50 Å to40,000 Å (pore diameter) to pore volume between 1,000 Å to 10,000 Å(pore diameter) of the sorbent is smaller than 2:1.
 16. The method ofclaim 15, wherein the toxin has a molecular weight in the range of from50,000 Daltons to 450,000 Daltons.
 17. The method of claim 1, whereinthe sorbent has a pore structure such that the total pore volume of poresize in the range of 50 Å to 40,000 Å is greater than 0.5 cc/g to 5.0cc/g dry sorbent; wherein the ratio of pore volume between 50 Å to40,000 Å (pore diameter) to pore volume between 10,000 Å to 40,000 Å(pore diameter) of the sorbent is smaller than 3:1.
 18. The method ofclaim 1, wherein the sorbent comprises a plurality of pores comprisingat least one crosslinking agent, at least one monomer, and at least onedispersing agent.
 19. The method of claim 18, wherein the dispersingagent is one or more of hydroxyethyl cellulose, hydroxypropyl cellulose,poly (hydroxyethyl methacrylate), poly (hydroxyethyl acrylate), poly(hydroxypropyl methacrylate), poly (hydroxypropyl acrylate), poly(dimethylaminoethyl methacrylate), poly (dimethylaminoethyl acrylate),poly (diethylaminoethyl methacrylate), poly (diethylaminoethylacrylate), poly(vinyl alcohol), poly (N-vinylpyrrolidinone), salts ofpoly (methacrylic acid), or salts of poly(acrylic acid).
 20. The methodof claim 18, wherein the crosslinking agent is one or more ofdivinylbenzene, trivinylbenzene, divinylnaphthalene,trivinylcyclohexane, divinylsulfone, trimethylolpropane trimethacrylate,trimethylolpropane dimethacrylate, trimethylolpropane triacrylate,trimethylolpropane diacrylate, pentaerythrital dimethacrylates,pentaerythrital trimethacrylates, pentaerythrital, tetramethacrylates,pentaerythritol diacrylates, pentaerythritol triacrylates,pentaerythritol tetraacrylates, dipentaerythritol dimethacrylates,dipentaerythritol trimethacrylates, dipentaerythritoltetramethacrylates, dipentaerythritol diacrylates, dipentaerythritoltriacrylates, dipentaerythritol tetraacrylates, or divinylformamide. 21.The method of claim 18, wherein monomer is one or more of divinylbenzeneand ethylvinylbezene, styrene, ethylstyrene, acrylonitrile, butylmethacrylate, octyl methacrylate, butyl acrylate, octyl acrylate, cetylmethacrylate, cetyl acrylate, ethyl methacrylate, ethyl acrylate,vinyltoluene, vinylnaphthalene, vinylbenzyl alcohol, vinylformamide,methyl methacrylate, methyl acrylate, trivinylbenzene,divinylnaphthalene, trivinylcyclohexane, divinylsulfone,trimethylolpropane trimethacrylate, trimethylolpropane dimethacrylate,trimethylolpropane triacrylate, trimethylolpropane diacrylate,pentaerythritol dimethacrylate, pentaerythritol trimethacrylate,pentaerythritol tetramethacrylate, pentaerythritol diacrylate,pentaerythritol triacrylate, pentaerythritol tetraacrylate,dipentaerythritol dimethacrylate, dipentaerythritol trimethacrylate,dipentaerythritol tetramethacrylate, dipentaerythritol diacrylate,dipentaerythritol triacrylate, dipentaerythritol tetraacrylate,divinylformamide and mixtures thereof.
 22. The method of claim 1,wherein the sorbent is a mixture of sorbents with two or more differentpore sizes.
 23. The method of claim 1, wherein the sorbent is formulatedas a powder, tablet, capsule, solution, gel tab, dispersion, slurry,suppository, or suspension.
 24. The method of claim 1, wherein thesorbent is admixed with food, fluid, or any combination thereof.
 25. Themethod of claim 1, wherein the toxin is one or both of Clostridiumdifficile Toxin A and Clostridium difficile Toxin B.